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Anti-Cyclin D1 antibody (ab7958)

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1 question for ab7958

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Question 1

Monday 08-November-2004

BATCH NUMBER 64979 ORDER NUMBER 300472

DESCRIPTION OF THE PROBLEM No signal

SAMPLE DLD1 human cells. Cell extract

PRIMARY ANTIBODY Your rabbit anti-cyclinD1 at 1 in 500 as recommended in TBS 5% non-fat milk. 1hour RT or overnight at 4oC. Wash 3x 5mins tBS 0.05% Tween20

SECONDARY ANTIBODY HRP goat anti rabbit (Santa Cruz) at 1 in 2500 in TBS 5% non-fat milk. 1hour RT. Wash 3x 5mins tBS 0.05% Tween20

DETECTION METHOD ECL

POSITIVE AND NEGATIVE CONTROLS USED Control is a parallel blot in exactly the same conditions but with a different primary antibody. This blot worked very well.

ANTIBODY STORAGE CONDITIONS 4oC as supplied

SAMPLE PREPARATION Laemlli buffer with protease inhibitors. Samples heated prioir to loading on gel

AMOUNT OF PROTEIN LOADED 20ug per lane

ELECTROPHORESIS/GEL CONDITIONS reducing. 10% gel

TRANSFER AND BLOCKING CONDITIONS Transfer 90 mins 35V. Block with TBS with 5% non-fat milk overnight

HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 4 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes

WHAT STEPS HAVE YOU ALTERED? Primary incubation time and temp.

ADDITIONAL NOTES Can I get a different antibody please! How about a mouse anti-Cyclin D1 e.g. ab6152

ANSWER:

  

Thank you for your enquiry and I'm sorry to hear that you are experiencing difficulty with this antibody. Did you run a positive control? We do recommend using Jurkat lysate as a positive control. You mentioned that you ran a parallel blot with a different primary antibody - was it a different cyclin D1 antibody or a completely different antibody?

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